VEGF-induced neoangiogenesis is mediated by NAADP and two-pore channel-2-dependent Ca2+ signaling
VEGF-induced neoangiogenesis is mediated by NAADP and two-pore channel-2-dependent Ca2+ signaling
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DOI：
10.1073/pnas.1406029111
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发表时间：
2014-11-04
影响因子：
11.1
通讯作者：
Filippini, Antonio
中科院分区：
文献类型：
--
作者：
Favia, Annarita;Desideri, Marianna;Filippini, Antonio
血管内皮生长因子（VEGF）及其受体VEGFR 1/VEGFR 2在控制血管生成，包括实体瘤的血管生成中起主要作用。在这里，我们描述了一个特定的Ca 2+信号通路连接到VEGFR 2受体亚型，控制内皮细胞（ECs）对VEGF的关键血管生成反应。该途径的关键步骤是参与有效的Ca 2+动员信使，烟酸腺嘌呤二核苷酸磷酸（NAADP），和酸性细胞内Ca 2+储存的双孔通道TPC 2亚型的特异性接合，导致Ca 2+释放和血管生成反应。发现使用NAADP拮抗剂Ned-19或使用Tpcn 2（-/-）小鼠遗传地靶向该细胞内途径在体外和体内抑制对VEGF的血管生成反应。在人脐静脉内皮细胞（HUVECs）中，Ned-19消除VEGF诱导的Ca 2+释放，损害ERK 1/2、Akt、eNOS、JNK的磷酸化、细胞增殖、细胞迁移和毛细血管样管形成。有趣的是，Tpcn 2 shRNA处理废除VEGF诱导的Ca 2+释放和毛细血管样管形成。重要的是，在小鼠体内VEGF诱导的血管形成基质胶塞被Ned-19消除，最值得注意的是，未能发生在Tpcn 2（-/-）小鼠，但不受影响的Tpcn 1（-/-）动物。这些结果表明VEGFR 2/NAADP/TPC 2/Ca 2+信号通路对于VEGF诱导的体外和体内血管生成是至关重要的。鉴于VEGF可以根据激活的信号转导途径的平衡引发促血管生成和抗血管生成反应，靶向特定的VEGFR 2下游信号转导途径可以改变这种平衡，可能导致更精细定制的治疗策略。
Vascular endothelial growth factor (VEGF) and its receptors VEGFR1/VEGFR2 play major roles in controlling angiogenesis, including vascularization of solid tumors. Here we describe a specific Ca2+ signaling pathway linked to the VEGFR2 receptor subtype, controlling the critical angiogenic responses of endothelial cells (ECs) to VEGF. Key steps of this pathway are the involvement of the potent Ca2+ mobilizing messenger, nicotinic acid adenine-dinucleotide phosphate (NAADP), and the specific engagement of the two-pore channel TPC2 subtype on acidic intracellular Ca2+ stores, resulting in Ca2+ release and angiogenic responses. Targeting this intracellular pathway pharmacologically using the NAADP antagonist Ned-19 or genetically using Tpcn2(-/-) mice was found to inhibit angiogenic responses to VEGF in vitro and in vivo. In human umbilical vein endothelial cells (HUVECs) Ned-19 abolished VEGF-induced Ca2+ release, impairing phosphorylation of ERK1/2, Akt, eNOS, JNK, cell proliferation, cell migration, and capillary-like tube formation. Interestingly, Tpcn2 shRNA treatment abolished VEGF-induced Ca2+ release and capillary-like tube formation. Importantly, in vivo VEGF-induced vessel formation in matrigel plugs in mice was abolished by Ned-19 and, most notably, failed to occur in Tpcn2(-/-) mice, but was unaffected in Tpcn1(-/-) animals. These results demonstrate that a VEGFR2/NAADP/TPC2/Ca2+ signaling pathway is critical for VEGF-induced angiogenesis in vitro and in vivo. Given that VEGF can elicit both pro- and antiangiogenic responses depending upon the balance of signal transduction pathways activated, targeting specific VEGFR2 downstream signaling pathways could modify this balance, potentially leading to more finely tailored therapeutic strategies.